ES2209877T3 - USE OF A DETERMINED HIALURONIDASE TO ELIMINATE THE SCARS, OPACITIES AND CORNEAL CLOUDS. - Google Patents

Abstract

The invention disclosed herein relates to biochemical methods for the elimination of corneal collagen fiber disorganization to improve vision. Disorganization of corneal collagen fibers is seen in corneal scars, corneal opacification and corneal haze. In addition, the invention relates to biochemical methods for the elimination of corneal collagen fiber disorganization resulting from accidental traumatic injury to the cornea and from refractive surgery for such as radial keratotomy (RK), photorefractive keratectomy (PRK), and laser in situ keratomileusis (LASIK) so as to improve visual acuity and quality of vision.

Description

Use of a given hyaluronidase for remove scars, opacities and clouds corneal

The invention presented below will be refers to biochemical means for scars removal, opacities and corneal clouds.

The cornea is the transparent dome located in the anterior part of the eyeball through which the light passes. Approximately eighty percent of the focusing ability, or Refraction, of the eye is located in the cornea. The loss of the corneal transparency, secondary to trauma or scarring, or any other cause of opacity, may result in reduced visual acuity and even blindness.

The overall prevalence of opaque eyes serious is estimated at more than three million, with more than 200,000 new additional cases per year. The current treatment of opacities corneal associated to significant defects of visual acuity It is a form of corneal transplant called keratoplasty penetrating or laminar (PKP, LKP), using donor tissue from cornea. Although, in general, this surgical technique is considered safe and effective, among the associated risks are the failure of the or graft rejection and infections transmitted through the donor cornea (for example, rabies, HIV, etc.) or the surgical procedure (for example, HIV, infections Staphylococcal, etc.). In addition to the various side effects, the number of corneal transplants that can be performed is limited by the number of corneas available for transplantation. Until now, the availability of donor corneal tissue adequate for the transplant does not cover the needs of The patient population.

Current methods to correct refractive defects of the eye, such as glasses, contact lenses, radial keratotomy, photorefractive keratotomy or laser in situ keratomileusis have not been useful for the removal of corneal opacities. Preliminary data suggest that radial keratotomy, photorefractive keratotomy or laser in situ keratomileusis can produce corneal clouds in some patients.

What is necessary for the treatment of The loss of corneal transparency is a biochemical method that Increase the transparency of the cornea. The methods and compositions presented below satisfy this need Long awaited.

The invention presented below relates to biochemical means for the elimination of the disorganization of corneal collagen fibers in order to improve vision. The disorganization of the collagen fibers is observed in the scars, opacities and corneal clouds. In addition, the invention is related to biochemical means for the elimination of the disorganization of the collagen fibers produced by traumatic injuries of the cornea, accidental or produced during refractive surgery such as radial keratotomy (RK), photorefractive keratotomy (PRK) and laser keratomileusis in situ (LASIK), performed to improve visual acuity and vision quality.

The invention presented below refers to to the improvement of visual transparency through the use of media biochemicals The biochemical means eliminate the scars, opacities and optical aberrations including clouds Corneal mammals, without corneal surgery, by Treatment of the cause of corneal distortion. The invention is useful to reduce corneal disorganization by modification of stromal proteins and proteoglycans corneal

The cornea is a transparent eye structure with multiple components through which the light passes and reaches Then the retina. The transparency of the cornea in the light is consequence of the exclusive extracellular matrix of the stroma corneal The stroma is a layer of tissue organized in sheets of collagen or densely packed parallel collagen fibers, embedded in a hydrated matrix of glycoproteins and Proteoglycans Size, regularity and precise spacing of the fibrillar structures are the physical characteristics essential for corneal transparency (Maurice, 1957).

The role of glycoproteins is not well known and corneal proteoglycans in the achievement and maintenance of corneal transparency It has been theorized that the proteoglycans of stroma have a role in regulating the spacing of collagen fibers (Hassell et al., 1983). Although not yet clear the precise role of proteoglycans, it is thought that influence the hydration, thickness and transparency of the cornea (Borcheding et al., 1975). It is still unknown functional significance of hyaluronic acid in the cornea, except during development (Toole and Trestad, 1971) and in some corneal alterations (Fitzsimmons et al., 1994).

In some opaque corneal scars it has since the scar contains collagen fibers with a diameter abnormally large and that there is an abnormal spacing interfibrillar (Schwarz and Keyserlingk, 1969). However, during healing of the corneal wounds produced in the rabbit, the initial opaque scars contain collagen fibers that they usually have a normal diameter and are irregularly spaced in the tissue. The diameter of collagen fibers does not changes significantly after a year of healing but the spacing between the fibrils returns to normal, with a concomitant decrease in the opacity of the scar (Cintron and Kublin, 1977 and Cintron et al., 1978).

A publication by Hassell et al., Of 1983, has shown that opaque scars containing wide spaces interfibrillar also used to contain chondroitin proteoglycans abnormally large sulfate with side chains of normal sized glycosaminoglycans. These dull scars they also lacked keratan sulfate proteoglycans but They contained hyaluronic acid. The biochemical analysis of proteoglycans from corneal scars in the rabbit, compared with those of the normal cornea adjacent to the scar shows that these areas synthesize demonstrably different proteoglycans each other (Cintron et al., 1990).

Hassell et al., (1980) analyzed samples corneals obtained during surgery of patients with dystrophy corneal macular These authors found that the cells of the normal corneas synthesized chondroitin sulfate proteoglycans and keratan sulfate similar to those present in monkey corneas and bovine Cells from the corneas with dystrophy macular synthesized a normal chondroitin sulfate proteoglycan but they did not synthesize keratan sulfate or proteoglycan from keratan mature sulfate On the contrary, the cells synthesized a glycoprotein with an abnormally oligosaccharide side chain long

As stated above, various corneal injuries lead to scarring and opacities corneal By itself, the opacity of the scar tissue is the result of the collagen fibers appeared during the process of healing lack the level of organization found in the corneal tissue not injured. Penetrating keratoplasty, which uses donor corneal tissue as a corneal graft, it is the only technique currently available for the elimination of corneal scars or opacities. The inconveniences associated with Penetrating keratoplasty are: (a) the availability of corneas of donors for surgery, (b) the compatibility of the cornea of the donor and probability of graft survival, (c) rejection of the cornea graft and (d) graft infection. The compositions and methods shown in the present invention offer a non-surgical alterative to penetrating keratoplasty.

Corneal transparency may result altered in a more subtle way than observed in previously mentioned injuries. In certain situations, the appearance of monochromatic optical aberrations can make decrease visual acuity (VA) of one eye. Based on the mosaic structures of the retina, visual acuity of the eye human could be 10/20 or higher: however, rarely Achieve this visual acuity. The two optical circumstances responsible for the suboptimal level of visual acuity are: diffraction due to pupil size and aberrations monochromatic (Campbell et al., 1974). The limitations of the visual acuity caused by diffraction decreases with increasing pupil size and can play an important role only in pupils less than 2 mm. However, the optical errors of a higher order (aberrations) of the human eye demonstrate a opposite behavior and may increase by increasing the diameter of The pupil.

The shape of the human cornea and lens It is designed naturally to minimize these aberrations. To our knowledge, the monochromatic aberrations of the human eye they have not been systematically studied in large series of individuals (Howland et al., 1977). Therefore, it lacks the average values in a standard population. However, the loss visual acuity by introducing optical aberrations it can be clinically significant with the advent of the corrective refractive surgery.

Refractive surgery for myopia and astigmatism, such as radial keratotomy (RK), photorefractive keratectomy (PRK) and laser in situ keratomileusis (LASIK), induce a non-physiological corneal shape, with a flattened central area and greater power towards the periphery. This form produces an increase in optical aberrations (Seiler T. et al., 1993) (Pallikaris I., 1998) and can lead to visual losses that are detected in circumstances of light shortage (Seiler Treatment, et al., 1994) and by a low contrast visual acuity test (Verdon W., 1996). These side effects of refractive corneal surgery have a potential for public health problems of dimensions still unknown.

Corneal wavefront aberrations after PRK and LASIK have been compared in a prospective randomized study of 22 patients with bilateral myopia treated with PRK in one eye and LASIK in the other eye (Oshika, T. et al., 199). Prior to surgery, simulated pupil dilation from 3 mm to 7 mm caused a five to six fold increase in total aberrations. After surgery, the same dilation produced a 25-32 fold increase in total aberrations in the PRK treated group and a 28-46 fold increase in total aberrations in the LASIK treated group. Both photorefractive keratectomy and laser in situ keratomileusis significantly increased total wavefront aberrations and the values did not return to preoperative normality in a 12-month follow-up period.

Studies have been conducted in rabbits after LASIK treatment to evaluate the healing process of corneal wounds after 1, 2 and 9 months of surgery. The periodic histopathological evaluation of animal corneas showed disorganized collagen fibers along the interface of the corneal flap even 9 months after LASIK surgery. These results show that the corneal aberrations and the LASIK surgery wound healing process they persisted 9 months after LASIK (Kato T., 1999). The invention that is presented provides means with which to overcome the effect Collateral optical aberration of modern surgical techniques refractive

Without restricting any mechanism of action In particular, it has been theorized that the various corneal aberrations resulting from RK, PRK, LASIK and other surgical procedures are a consequence of the disorganization of the corneal collagen produced during the healing process. For example, after LASIK procedure, after placing the flap to cover the surgical procedure site, collagen will form to seal the incision. When this collagen is formed, it is thought to be distributed in a conformation that, in one degree or another, is less organized that the collagen of the areas not affected by the surgery. The reorganization of this material would lead to reduction of optical aberrations resulting from such types of surgery.

Misra, SS et al (1982), Indian Journal of Animal Sciences ( 52) 5 : 360-361, has shown that hyaluronidase administered subconjunctivally, at doses of 750 IU clarified acute and chronic corneal opacities in six animals.

The invention presented is a useful biochemical means for the elimination of corneal aberrations and disorganization of corneal collagen fibers resulting from traumatic corneal lesions or by refractive surgery for myopia, farsightedness and astigmatism, such as radial keratotomy ( RK), photorefractive keratectomy (PRK) and laser in situ keratomileusis (LASIK), and thus improve visual acuity and vision quality.

Accordingly, the present invention provides the use of hyaluronidase devoid of fractions of molecular weight greater than 100,000, between 60,000 and 70,000 and less of 40,000, using 10% SDS-PAGE, for manufacture of a medicine or composition to eliminate scars, opacities and clouds.

Hyaluronidase can be administered at a rate from 1 to 8,000 International Units.

The medication can be formulated to:

to)

injection intrastromal,

b)

application topical

c)

administration through contact lenses,

d)

administration single or

and)

administration to less twice

The aforementioned corneal opacity may result from a surgical procedure selected from the group formed by cataract surgery, corneal transplantation, radial keratotomy (RK), photorefractive keratectomy (PRK) and laser in situ keratomileusis ( LASIK).

According to the use of the invention, the introduction of the particular composition in the cornea of a individual improves visual acuity. The medications contemplated for use they can comprise a certain number of enzymes or others agents that give rise to the reorganization of collagen fibers corneal The enzymes that act are of particular interest altering, digesting or degrading in various ways components related or associated with collagen fibers corneal

Medications for use according to this invention may contain other chemicals that can be administer to reorganize corneal collagen fibers through various mechanisms of action. For example, a quantity of a specific reorganizing enzyme of the corneal fibers, to contacting the corneal stroma will accelerate the degradation rate of the corneal proteoglycans and will lead to the reorganization of the corneal collagen The resulting reorganization will clarify the scars, opacities and corneal clouds.

Gucosaminoglucanase enzymes present all them, the property of providing a clarification effect of corneal opacities as well as ability to eliminate aberrations and disorganization of collagen fibers corneal Among the specific enzymes are hyaluronidase, the keratinase, chondroitinase AC, chondroitinase B, chondroitinase ABC and the Chondroitin-4-sulfatase.

Metalloproteinases have an effect clarifier of corneal opacities and ability to eliminate aberrations and disorganization of collagen fibers corneal Specific examples of appropriate enzymes are the matrix metalloproteinase -1, matrix metalloproteinase -2, matrix metalloproteinase -3 and metalloproteinase of the matrix -9.

Protein kinase enzymes have an effect clarifier of corneal opacities and also capacity for elimination of aberrations and disorganization of fibers corneal collagen Specific examples of such enzymes are the Streptokinase and urokinase.

There are other enzymes that alter or modify the fibrillar structure of the corneal collagen and can be used in the invention. Appropriate enzymes induce modifications of the corneal collagen or its components and decrease opacities and the corneal optical aberrations.

Formulations

The invention presented contemplates the preparation of formulations that are effective for reorganization of corneal collagen. An aspect of the invention contemplates the formulation of an injectable solution containing a  reorganizing enzyme of the corneal collagen. The solution for injection It may contain certain inactive ingredients that make it substantially isotonic and provide an adequate pH for the injection into the eye Such an injectable solution may be initially lyophilized and subsequently reconstituted before its use.

A general formulation is presented in Table I injection of a thimerosal-free hyaluronidase preparation of this invention:

TABLE I

General formulation Ingredient Quantity Hyaluronidase / Hyaluronidase (ACS) up to 8,000 IU Lactose, USP / sorbitol 13.3 mg - 130.0 mg Phosphate, USP 5-200 mmoles

This ingredient formulation is prepared initially dissolving the ingredients in sterile water and filtering the solution under sterile conditions and is dispensed as solution or as lyophilisate until drying. The composition lyophilized can be packaged for further reconstitution before its use by adding a balanced saline solution or with Isotonic sterile saline. Such a balanced salt solution  typically contains 0.64% sodium chloride, 0.075% chloride potassium, 0.048% calcium chloride dihydrate, 0.03% sodium acetate trihydrate, 0.17% sodium citrate dihydrate, sodium hydroxide / hydrochloric acid to adjust the pH and water for injection up to 100%.

The term "hyaluronidase ACS" is used here to describe a preferred hyaluronidase, devoid of hyaluronidase fractions of molecular weight greater than 100,000, between 60,000 and 70,000 and below 40,000 (10% SDS-PAGE). Said hyaluronidase can proceed from the sheep testis and can be requested from Biozyme Biochemicals, 9939 Hilbert Street, San Diego, California 92131-1029. Applicants have determined that this specific distribution of the molecular weight of hyaluronidase ACS produces less ophthalmic toxicity than other preparations of hyaluronidase, maintaining the desired therapeutic efficacy in a series of ophthalmic applications.

The specific molecular weight distribution and the specific profile of the hyaluronidase enzyme activity (ACS) of the present invention, thimerosal exclusion from its formulation, or both factors, provide a preparation of hyaluronidase that is not toxic to the eye administered in doses in which other hyaluronidase preparations preserved with Thimerosal would produce toxic effects.

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TABLE II

Formulation specific Ingredient Quantity Hyaluronidase / Hyaluronidase (ACS) 1IU-8,000 IU Lactose USP / Sorbitol 13.3 mg - 150 mg USP Phosphate 5 mmol - 50 mmoles USP Sodium Chloride Until the isotonicity

TABLE III

Formulation specific Ingredient Quantity Hyaluronidase / Hyaluronidase (ACS) 1IU-50,000 IU Lactose USP / Sorbitol 13.3 mg - 150 mg USP Phosphate 5 mmol - 50 mmoles USP Sodium Chloride Until the isotonicity Sterile water for injection USP QS 2.0 ml (Concentration of hyaluronidase 500 IU / 20 µl)

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TABLE IV

Formulation specific Ingredient Quantity Hyaluronidase / Hyaluronidase (ACS) 2,500 IU Lactose USP / Sorbitol 10 mg USP Phosphate 5 mmoles USP Sodium Chloride Until the isotonicity Sterile water for injection USP CSP 2.0 ml (Concentration of hyaluronidase 25 IU / 20 µl)

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TABLE V

Formulation specific Ingredient Quantity Hyaluronidase / Hyaluronidase (ACS) 100 IU Lactose USP / Sorbitol 10 mg USP Phosphate 5 mmoles USP Sodium Chloride Until the isotonicity Sterile water for injection USP CSP 2.0 ml (Concentration of hyaluronidase 1 IU / 20 µl).

As described in the following examples, the ACS hyaluronidase specific formulation expressed in the Tables II-V (above) can be administered by injection, topical or contact lens application, in doses sufficient to achieve the desirable therapeutic effect, including, but without limitation, the elimination of corneal opacities, without cause no significant toxicity to the eye.

Determination of enzymes and agents appropriate for reorganization of corneal collagen and dosage

The following is based on technical knowledge previous.

The reorganizing chemicals of corneal collagen, such as various agents and enzymes used in the methods of the present invention, in addition to the doses appropriate agents and enzymes can be determined by An expert professional through routine experimentation. Bliss experimentation can comprise the analysis of a dose of a enzyme or agent in donor balloons (eyes) mounted on plastic concavities or testing said dose in animals of laboratory. Briefly, to determine if an enzyme or agent is effective for the reorganization of the collagen of a cornea without to produce toxicity, said enzyme or agent is first mixed with a Pharmaceutically acceptable vehicle for a mammal. Preferably the enzyme or agent should be lyophilized (in the form of dry powder) and dissolves in isotonic saline. However, a expert professional knows that a number of vehicles can be used pharmacologically acceptable that do not interfere with the operation of an enzyme or agent. Such a procedure is also continue to analyze different doses of a compound of interest.

Next, a cornea is given a test dose of the enzyme or agent to analyze its effects on the reorganization of collagen and toxic effects. In a procedure of analysis of the candidates, the enzyme or agent to Test is first administered to the corneas. This procedure is particularly preferable for determining the effect of an agent or enzyme on the human cornea since this way you can analyze the effect on it without subjecting a living person to Experimentation

The test dose of the enzyme or agent is administer to the donor cornea. Such administration can, by example, be done by injecting the enzyme into the cornea. Normally the lens will be opacified after this step due to the introduction of water into the eye and the change in the index of refraction of the eye. After a trial period, the cornea mounted to determine if any Collagen reorganization, and if so, see the degree of it. The toxicity resulting from the injection is also determined.

The cornea exam can be done, by example, using the slit lamp, to determine the transparency; by pachymetry to determine the thickness of the cornea; by measuring the resistance of the cornea to the traction; by measuring compliance and with keratometry to measure the central corneal curvature. The values determined in these tests are compared with the values determined before administration of the agent or enzyme.

In addition, a cornea treated in an eyeball mounted can be subjected to a series of additional tests to determine the strength and viability of the cornea after treatment. For example, optical microscopy can be performed, scanning electron microscopy, lightning diffraction analysis X and transmission electron microscopy to study the corneal morphology; tissue culture can be useful for determine the viability of corneal cells after treatment; the biochemical study of the collagen and other structural components of the cornea after treatment.

Previous tests on eyeballs and Donor corneas are used to verify that the use of a particular enzyme or agent increases the transparency of the cornea without decreasing the viability of corneal cells or damage the structural integrity of the cornea. Nevertheless, It is also desirable to test the enzyme or agent in corneas of experimental animals to ensure that the candidate agent does not has unexpected effects on living mammals that have not been discovered during tests on donor eyes. With the In order to analyze the effect of a particular enzyme or agent, it administer a dose in a pharmacologically acceptable vehicle to an experimental animal, in this case a mammal, so that The agent reaches the cornea of the animal.

After the administration of an agent in the cornea of the animal, it can be subjected to the following studies: slit lamp examination to determine transparency of the cornea, anterior chamber and iris, pachymetry to measure corneal thickness, computerized corneal topography to evaluate topographic changes of the corneal surface, measure of the elasticity of the cornea, tonometry to measure pressure intraocular, fundoscopic examination to evaluate the optic nerve and the retina, keratometry to measure the central corneal curvature, retinoscopy to measure refractive defect, staining with fluorescein or Bengal rose to identify lesions of the corneal epithelium and indirect ophthalmoscopy. The values determined in these explorations can be compared with values determined before administration of the enzyme or agent and also with the values determined in the untreated eye of the animal

In addition, the treated cornea of the animal can be subjected to another series of tests to determine transparency, resistance and viability after treatment. For example, you can perform optical microscopy and transmission electron microscopy and scanning, to study the morphology of the cornea; the cultivation of tissues can be useful to determine the viability of corneal cells after treatment; you can also perform the biochemical study of collagen and other structural components of the cornea after treatment.

Other enzymes and modifying agents of corneal collagen not mentioned here and the appropriate doses of such Enzymes, known or unknown, can be determined as described above in relation to the determined enzymes and the corresponding doses.

Administration Methods

The applicant has identified the use of a hyaluronidase devoid of higher molecular weight fractions to 100,000, between 60,000 and 70,000, and less than 40,000 using 10% SDS-PAGE, for the manufacture of a non-surgical clearing medication scars and corneal opacities. The contact of the cornea with the medicament prepared according to the invention is effective to accelerate obtaining transparency of corneal opacities. The Clarification of corneal opacity can be achieved without any other surgical manipulation or corneal resection, with which avoid the risks and potential complications associated with penetrating keratoplasty

The medications (compositions) mentioned They can be administered by any of the known routes. By example, in one application, the composition is injected directly in the eye, in a location proximal to the cornea. In this way of application, the composition includes a pharmacologically vehicle  acceptable that does not alter the effectiveness of it.

In the rest of the description that follows, the reference to "corneal collagen reorganizing enzyme" refers to hyaluronidase of the medicaments of the invention.

In a particular embodiment of the present invention, the route of administration of the reorganizing enzyme of the Corneal collagen is intrastromal injection. In this application the corneal collagen reorganizing enzyme is injected with a needle, directly in the stroma located in the anterior part Of the eye.

In another application of the present invention, the corneal collagen reorganizing enzyme is administered topically, in the form of eye drops. The number of drops should be sufficient to administer the desired concentration of the enzyme to the cornea of the individual. The method of administration by eye drops may be superior to the injection because produces less corneal discomfort to the patient.

In another application, media can be used alternatives to facilitate diffusion in the eye to the cornea. These means include the use of liposomes for the delivery of The enzyme or agent. The enzyme is packaged in liposomes that can cross the liposoluble membrane of the corneal epithelium and stroma corneal Another way to facilitate dissemination is the use of a electric current to make the outer membrane more permeable from the eye to the passage of enzymes, that is, by iontophoresis

In another application the iontophoresis is used to administer the reorganizing reagents of the corneal collagen. Using this procedure, an electric current that passes to through a saline solution causes the agent to pass into the eye as charged particles.

In another additional application of this invention, the corneal collagen reorganizing enzyme is administered through contact lenses. As an example of a application of this invention, is loaded into a chamber within a rigid contact lens, preferably gas permeable, a amount of the corneal collagen reorganizing enzyme. Alternatively, the enzyme can be charged or impregnated with a lens. soft able to capture the enzyme or agent soaking the soft lens of a solution thereof. The enzyme can also be loaded into a series of soft and rigid lenses.

In all the following additions of a contact lens for administration of reorganizing enzyme of corneal collagen, it is given as it diffuses (or is released) from the lens chamber or from the material of the lens (if the enzyme has been embedded in a soft lens). The doses of different vehicles with contact lenses can be optimized by the professional expert through the routine experimentation

According to a method of administration by lenses contact, collagen reorganizing enzymes and agents Corneal can be applied to the eye using contact lenses rigid. These lenses can be made of Known fluorosilicone acrylate, gas permeable. The lens is supplied with an internal camera for storage of the enzyme or reorganizing agent of corneal collagen. Preferably, the chamber will contain a space radially symmetrical surrounding the entire lens, located between the surface front and back of it.

For the present purpose, the rigid lens can be made turning, modeling or milling a posterior component and an earlier component from a lens disc that, during manufacturing, can be fixed together to form a lens unit using the bonding or bonding techniques known to The experts. The camera can be made by turning an annular recess on the convex surface of the posterior component of the lens before of the end of its manufacture. You can use any of the varieties of size and a preferred form is the one that provides an annular chamber of an approximate width of 1.0 mm to 1.5 mm and a depth between 0.05 and 0.10 mm.

On the back of the lens there will be multiple microscopic holes that allow communication between the chamber and eye and facilitate the delayed release of the enzyme reorganizing corneal collagen to the cornea. Holes they can be done by mechanical or laser drilling or by molding before the assembly of the previous components and rear of the lens In one application the holes are made using a mechanical drill with a microcarbon tip.

The pumping action of the eyelids, combined With natural tearing, it helps release the enzyme reorganizing the corneal collagen through the tiny holes Preferably, the holes are made by mechanical drilling with a microcarbon tip and will have a diameter between 0.002 and 0.010 mm and preferably about 0.005 mm. The number and diameter of the holes may vary for modify the characteristics of the release pattern, such as It is evident to the expert professional. However, in In general, for the specified range of diameters the use is contemplated 3 to 10 holes.

In a lens application, the back of it has a central thickness of about 0.12 mm and a recess annular with a depth of about 0.075 mm. To provide the communication with the rear surface of the lens is perforated certain number of holes, each with a diameter of about 0.005 mm, evenly spaced, on the periphery of the camera. The number of lens holes will vary depending on the desired rate of administration of enzyme or reorganizing agent of corneal collagen from the chamber.

The front part of the lens, with a thickness center of about 0.12 mm is subsequently fixed to the back closing the annular recess and forming a chamber, so that it obtains a lens with a total central thickness of about 0.24 mm. The binding can be done by applying a small amount of agent bonding, such as the 3M Concise ™ enamel system (St. Paul, Minnesota). Expert professionals know other means of junction of the anterior and posterior parts of the lens Contact.

In another aspect a lens of contact consisting of two layers laminated together. In this Advantageous contact lens design can be manufactured cameras larger size for storage of the reorganizing enzyme of corneal collagen.

In this contact lens, a anterior part with an anterior surface and a surface later. You can also manufacture a back of the lens with an anterior and a posterior surface. Perimeter external of the posterior surface of the anterior part can designed so that it has the same radius of curvature as the outer perimeter of the anterior surface of the back. In this way, when the back surface of the anterior part and the front surface of the back are laminated together, the sealing between the external perimeters occurs of the anterior and posterior parts.

However, in a central part of the part anterior, the posterior surface may have a radius of curvature more pronounced than the anterior surface of a central part of the back part. As a result of this greater pronouncement of the radius curvature, when the anterior part and the central part from the back they are laminated together, a camera is formed between the central part of the previous part and the central part of the back of the contact lens. Camera volume can be adjusted by changing the radius of surface curvature posterior of the central part and of the anterior surface of the central part, which is evident to the professional expert.

In this design, before manufacturing, it make one or more holes in the central part of the part back of the contact lens. The holes can be made by mechanical drilling with a microcarbon tip or with laser, for example, with argon laser, and will have a diameter approximately 0.002 mm to 0.010 mm, preferably about 0.005 mm The number and diameter of the holes may vary to vary the characteristics of the release pattern, which results Obvious to the professional expert. Therefore, the rate of dispensing a dose of collagen reorganizing enzyme corneal from the chamber is largely controlled by size and the number of holes present in the central part of the part rear of the lens However, in general, with the range of diameters specified above, it is contemplated to use 3 to 10 holes These holes will be spaced around the part center of the back of the contact lens for provide communication between the camera and the surface of the eye on the individual wearing the lens.

In a preferred application of this lens, the back of it can have a central thickness of about 0.125 mm The anterior part may have a central thickness of about 0.125 mm When the front and back are joined, the lens created has a total central thickness of about 0.24 mm. The Union it can be done by applying a sufficient amount of an agent of bonding, such as the 3M Concise ™ enamel system (St. Paul, Minnesota).

In an alternative embodiment, it is provided a contact lens that has a peripheral camera instead of the camera in the central part of it. In this embodiment the lens can be composed of an anterior part and a part back laminated together. In this application, the camera will Locate in the middle part of the lens.

In another embodiment, the camera can be formed in the middle part of the lens providing an area of the back surface of the front part of the lens that has a radius of curvature more pronounced than that of the rest of the back surface of the front part of the lens. Like in the previous application of contact lens with camera, the volume of the corneal collagen reorganizing enzyme that may contain the lens, and therefore, which is administered to the patient, is in great measure determined by the radius of surface curvature back of the inside of the lens in the middle of it, as well as the radius of curvature of the surface front of the back of the lens in its part intermediate.

The back of the lens is also provided with holes in the back, in the part intermediate lens. These holes serve to allow transfer of the contents of the chamber to the patient's eye. The number and size of the holes will determine, to a large extent, the rate of corneal collagen reorganizing enzyme supplied to the eye.

Although it has been described that the applications of a contact lens provided with camera are achieved by the joint lamination of the front and back of the lens, expert professionals know other methods that enable manufacture of the cameras described.

According to a method of additional administration of the invention presented, a lens can be impregnated or charged therapeutic or soft protection with a dose of enzyme reorganizing corneal collagen. The soft lens can then adjust to the cornea and keep it for a few hours to release the enzyme or agent to it. When the enzyme or agent have sufficiently modified the corneal collagen, the lens soft dissolves or withdraws.

Other types of soft lens materials tend to capture less the solution that contains an enzyme or agent and also to release it faster. Among the examples of such materials are the common hydrophilic materials for slow soft, such as etafilcon A and phemfilcon A, which can be get on Vistacon and Wesley Jessen. These lenses can be disposable or long lasting. The lenses with a content of H2O between approximately 58% and 70% may result useful in the method shown.

Fields of use

One of the contemplated uses of this Invention is the treatment of corneal opacities. The corneal opacities may be due to age, as the corneal collagen is losing its organization, or being a consequence of some trauma. The invention is useful for the elimination of such corneal opacities.

Another contemplated use of the present invention is the rehabilitation of irregularities and the improvement of refractive defects that result from various forms of surgery corneal The surgical procedures referred to include the photorefractive keratectomy (PRK), LASIK, keratotomy radial (RK), corneal transplantation and cataract surgery. For example, LASIK is having a procedure. Extremely frequent worldwide. Although by itself the technique is very successful, incidents and effects are increasing collateral treatment attributable to disorganization of corneal collagen The present invention can be used for improve the visual acuity of patients after remodeling Corneal surgery using various procedures.

In this application of the invention, would identify the patient undergoing the LASIK procedure and would select an acceptable collagen reorganizing agent corneal Administration of the collagen reorganizing enzyme Corneal can be performed before, during or after the procedure surgical. In one application, the patient can receive the enzyme corneal collagen reorganizer after the procedure surgical, to facilitate proper healing of the cornea. How It has been said previously, the routes of administration contemplated They include injection and topical administration.

Similarly, the present invention also can improve the chances of success of other procedures corneal, such as corneal transplant surgery and surgery The falls One of the most frequent reasons for the failure of surgical procedures such as corneal transplantation is the refractive defects after surgery otherwise successful The enzyme presented here can be used to correct the refractive defect that appeared as a result of the disease, surgery or other conditions. Besides, the invention, by improving visual acuity resulting from a surgical procedure will reduce or eliminate the need for subsequent surgical procedures that are necessary today to eliminate undesirable refractive defects.

To use the invention in order to obtain These additional clinical benefits must first be identified to patients with corneal opacities, disorganization of corneal collagen or undergoing corneal manipulation. Such identification is usually done by an ophthalmologist or other expert professional who can identify corneal opacity, the disorganization of corneal collagen or corneal manipulation. TO then the administration methods will be used previously described to induce the reorganization of corneal collagen.

Examples Example I Ophthalmic toxicity of thimerosal, hyaluronidase (ACS) e hyaluronidase (Wydase®) in the rabbit

Fifty-two (52) healthy rabbits of the New Zealand Cross variety (26 males and 26 females) of 1.5 to 2.5 kg in weight were individually marked and distributed in suspended individual cages. The animals were fed daily with food sold in the form of pellets and available tap water
ad libitum .

The animals were divided into thirteen groups of 4 animals each (2 males and 2 females). Two were selected animals (1 male and 1 female) from each group for photography of the Fundus and fluorescein angiography before treatment.

The background photograph was made subject to animal and visualizing the optic nerve, retinal arches and the fundus with a KOWA® RC-3 Fundus Camera Loaded with Kodak Gold 200 ASA film.

Fluorescein angiography was performed. injecting 1.5 ml of 2% sterile fluorescein solution via the marginal vein of the ear. Approximately 30 seconds after fluorescein injection was visible in the optic nerve, vessels retinians and fundus.

The next day the animals were anesthetized by intravenous administration of a combination of Ketamine Hydrochloride, 34 mg / kg, and xylazine, 5 mg / kg. The Eyelid retraction was performed with a palpebral speculum and the eyes were disinfected by washing with povidone iodine.

Experimental treatments with solution balanced saline (BSS), BSS + thimerosal, (Wydase®) or hyaluronidase (ACS) were administered by injection using a syringe tuberculin and a 30-gauge, 0.5-inch needle length. The hyaluronidase (ACS) solution used in this example was thimerosal free and constitutes the formulation Preferred hyaluronidase (ACS) of Table II exposed previously. Experimental treatments administered to each group of animals were the following:

TABLE VI

# Group Treatment one BSS two BSS + 0.0075 mg Thimerosal 3 BSS + 0.0025 mg Thimerosal 4 Hyaluronidase (Wydase) 1 IU 5 Hyaluronidase (Wydase) 15 IU 6 Hyaluronidase (Wydase) 30 IU 7 Hyaluronidase (Wydase) 50 IU 8 Hyaluronidase (Wydase) 150 IU 9 Hyaluronidase (ACS) 1 IU 10 Hyaluronidase (ACS) 15 IU eleven Hyaluronidase (ACS) 30 IU 12 Hyaluronidase (ACS) 50 IU 13 Hyaluronidase (ACS) 150 IU

The day after the injections (day 1) controlled the 26 animals that had undergone the Fundus photography and fluorescein angiography, using the same methods as in the pre-administration examination of the dose.

On day 2 after the injections, it they sacrificed using a drug based on sodium pentobarbital the 13 male rabbits selected for fundus photography and fluorescein angiography before dose administration and on day 1 as well as the 13 females not selected for Photography. The eyes were surgically removed and placed in a 2.5% glutaraldehyde fixative solution with buffer phosphate saline with a pH of 7.37. Alternatively, one of the Rabbits, chosen at random, were sacrificed with an injection of pentobarbital and was fixed by intracardiac injection of glutaraldehyde solution in the left ventricle to determine the effect of the fixing procedure on the histological findings of enucleated eyes.

On day 7, the 13 females that were previously had done them photography and fluorescein angiography were subjected to the same studies, following the methods previously described.

The remaining 26 animals were euthanized, as described above, 7 days after administration of the dose. They fixed their eyes in the same way described to those set on day 2. In addition, a randomly selected rabbit was undergoing the same fixation procedure by injection intracardiac glutaraldehyde described above for the animal previously selected at random.

The eyes of the animals treated in this example were examined macroscopically and microscopically in order to Detect treatment related toxicity. In Table VII a summary of the histological data indicative of toxicity or absence of toxicity in each treatment group.

In short, the eyes of the treated control group with BSS they showed no signs of toxicity 2 and 7 days after the dose.

The eyes of the animals of group 2, treated with BSS + thimerosal (0.0075 mg) showed no toxicity on day 2 but showed signs of hematorretinal barrier alteration at 7 days

Animals of group 3, treated with BSS + Thimerosal (0.025 mg) showed intense effects related to treatment on days 2 and 7 after administration of the dose.

Group 4 animals treated with Wydase® in doses of 1 IU they lacked signs of toxicity on days 2 and 7 but the eyes of the animals of the Nos groups. 5-8, treated with Wydase® in doses that ranged between 15 and 150 IU generally showed toxic effects related to the dose at 2 and 7 days of it.

The eyes of the animals of the groups of treatment Nos. 9-13, treated with hyaluronidase (ACS) in doses ranging from 1 to 150 IU lacked signs that indicate toxic effects on days 2 and 7 after dose.

Consequently, it was concluded that thimerosal and Wydase® formulation with thimerosal, in the doses tested, produce toxic effects on the eyes of rabbits and that the hyaluronidase (ACS) does not produce toxic effects in these animals, in the doses administered.

Table VII summarizes the results of the studies carried out on day 7. As shown in said Table VII, significant toxic effects were observed on day 7 in the eyes of rabbits treated with BSS plus thimerosal (0.0075 mg) e hyaluronidase (Wydase®) with all doses between 1 and 150 IU. By On the contrary, no toxic effects were observed in the treated eyes with hyaluronidase (ACS) in doses between 1 and 150 IU.

TABLE VII Toxic effects of a single intravitreal dose of BSS, BSS + thimerosal, hyaluronidase (ACS) and hyaluronidase (Wydase®) in the rabbit

one

Example II Ophthalmic toxicity of thimerosal, hyaluronidase and hyaluronidase (Wydase®) injected into the rabbit's cornea

Twenty (20) healthy rabbits of the New Zealand Cross variety, 1.5 to 2.5 kg in weight, were individually marked and placed in individual suspended cages. The animals were fed commercial feed in pellets and drank tap water ad libitum .

The animals were divided into 4 groups, each of 5 animals The 20 animals were examined by biomicroscopy with slit lamp and fluorescein staining to verify the condition of the cornea before treatment.

The next day the animals were anesthetized by intravenous injection of a combination of 34 mg / kg of Ketamine Hydrochloride and 5 mg Xylazine. They withdrew eyelids with a palpebral speculum and the eyes were disinfected by washing with povidone iodine.

Experimental treatments consisting of buffered saline solution, hyaluronidase were administered
(Wydase®) or hyaluronidase (ACS), by injection, using a 0.3 ml tuberculin syringe with a 0.5-inch 29-gauge needle. The hyaluronidase (ACS) used in this example was free of thimerosal and was the preferred specific hyaluronidase (ACS) formulation, previously described.

Experimental treatments administered to Each animal were the following:

GROUP no. TREATMENT Right eye Left eye one BSS Control no treaty two Hyaluronidase (Wydase) 25 IU Hyaluronidase (Wydase) 100 IU 3 Hyaluronidase (ACS) 25 IU Hyaluronidase (ACS) 100 IU 4 Hyaluronidase (ACS) 500 IU Hyaluronidase (ACS) 1,000 IU

On days 1, 7, 15 and 30 after the injection the eyes of the animals were examined macroscopically and by Biomicroscopy to check for signs of toxicity related to treatment.

In short, the eyes of animals treated with BSS and those of the untreated control group were exempt from toxicity.

The eyes of group 2 animals, treated with Hyaluronidase (Wydase®) preserved with thimerosal presented toxic lesions The eyes of the animals of groups 3 and 4, treated with hyaluronidase (ACS) showed no signs of toxicity.

Consequently, it was concluded that the formulation Wydase®, which contains thimerosal, produces toxic effects on the eyes of rabbits, in the doses tested. However, the hyaluronidase (ACS) does not cause toxic effects in these animals in The doses tested.

Example III Efficacy of hyaluronidase in the clarification of corneal opacities in the corneas of human donors

Corneas were obtained from human donors with stroma scars, between 24 and 48 hours after death. The corneas were photographed to document the position of the scar. They were then placed in organ culture for 72 hours The culture medium consisted of modified MEM of Dulbecco, serum free supplemented with chondroitin sulfate, EGF, dextran, selenium and vitamin A. The corneas were grown in 10 mm sterile culture plates containing 10 to 12 ml of the culture medium, with the epithelial surface facing up. The plate placed on a swing platform in a humidified incubator at 37º C and in an environment with 5% CO2, making it oscillate so that the epithelial tissue was exposed so intermittent to an air / liquid interface.

On day 3 hyaluronidase (ACS) was injected reconstituted with saline (500 IU / 20 µl) in the stroma corneal adjacent to the scar, with the needle bevel directed towards the center of the cornea. The cornea control group is injected with saline on day 3. All corneas returned to place in the culture medium. The problem corneas were injected subsequently with hyaluronidase (ACS) (500 IU / 20 µl) in the day 7, as previously described.

The corneas were examined daily for Control the resolution of scars. The resolution is determined based on the presence or absence of opacity corneal At the conclusion of the experiment, the photographs were photographed again. corneas and were processed for histological and microscopy study electronics.

Results

The corneas showed a fine morphology preserved with minimal swelling by the crop. The corneas are cultured for 72 hours before hyaluronidase injection (ACS) to ensure that the crop, by itself, had no effect about the morphology of the scars. The results of this experiment is summarized in the following table. They were not observed Scar changes during this period. Immediately after of hyaluronidase (ACS) injection in the corneal stroma is develops a cloud area at the injection site. I know think that this cloud is due to the injection of the liquid into the stroma and typically resolves spontaneously at 4-8 hours of injection. To date, they have studied a total of 14 corneas and in 12 of the 14, within a period of 2 to 4 days after treatment with hyaluronidase (ACS), the scars were no longer visible on inspection ocular. The scars were variable in terms of type and location, from post-surgical peripheral scars, as frequently observed after insertion of intraocular lenses IOL, to scars secondary to accidental trauma. The Most of the scars examined were of unknown cause.

Intrastromal injection of hyaluronidase (ACS) It turns out to be very effective, as an alternative to corneal transplantation to the resolution of corneal opacities.

two

3

Example IV Effect of hyaluronidase on the structure of the corneas of human donors

For the following example corneas of human donors obtained between 24 and 48 hours after death from the donor First, the corneas were dissected and fixed in 1% glutaraldehyde solution, followed by processing and imbibition in plastic, or placed in culture medium. The Corneas subjected to culture were grouped as follows: a) untreated controls; b) controls injected with saline, and c) treatment group injected with hyaluronidase (500 IU / 20 µl). The corneas were cultured with the epithelial side toward top, in sterile 10 mm culture plates, containing 10 a 12 ml of culture medium. The culture medium consisted of MEM modified from Dulbecco, serum free, supplemented with sulfate of chondroitin, EGF, dextran, selenium and vitamin A. Plaque is placed on a rocking platform in a humidified incubator, to 37º C and in an environment with 5% CO2, making it oscillate to that the epithelial surface was exposed so intermittent to an air / liquid interface.

\hbox{Tabla VIII.}

At the end of the treatment period the corneas were dissected and fixed in 1% glutaraldehyde solution, processed and imbibed in plastic resin. Complete cross sections (1 µm) of the central stromal regions were made, stained and examined by optical microscopy. In addition, subsequent ultra-thin cuts (0.1 µm) were made and studied by electron microscopy. The ultrastructural changes in the density of collagen fibers were determined in electronic microphotographs (at 124,000 magnifications) counting the average of collagen fibers per 180 square millimeters. The results of this study are summarized in the

 \ hbox {Table VIII.} 

TABLE VIII Density of collagen fibers in electronic photomicrographs (increases: 124,000 X)

4

Results

The results show an increase of 18.62% in the reorganization of collagen fibers 48 hours after hyaluronidase injection. This magnitude of density and corneal reorganization of an 80 year old woman is statistically very significant compared to the cornea no treated of the 65 year old male. Intrastromal injection of hyaluronidase (ACS) turned out to be very effective for reorganization of corneal collagen fibers.

Example V Efficacy of hyaluronidase for clarification of the cornea in humans

In this study two (2) patients were treated with corneal opacities with a single hyaluronidase injection intrastromal, in doses of 500 IU patient # 1 and 800 IU the patient # 2. The results of this experiment are summarized in the Table IX

TABLE IX

Patient# Dose from Opacity Opacity Opacity Opacity hyaluronidase corneal corneal corneal corneal before of the 1 week 1 month 6 months treatment post treatment post treatment post treatment Woman 500 IU Dense Molecule Cornea Cornea opacity corneal transparent transparent corneal + one Male 800 IU Scar Opacity Cornea *Cornea corneal corneal previous transparent transparent * Two months after treatment

In the two (2) patients treated in this example, the corneal opacity became transparent enough within a period of 30 days after a single injection of hyaluronidase (ACS). Such corneal clarification would not have been produced in patients without hyaluronidase treatment.

Example VI Use of hyaluronidase for the resolution of scars corneal

In a patient who presents with a scar corneal identifies the need for collagen reorganization corneal to improve your visual acuity. The patient is treated with intrastromal injection of hyaluronidase ACS as shown in the Table IV The pharmaceutical composition is injected into the stroma of the eye with the corneal scar.

ACS hyaluronidase treatment of the Present invention starts after a complete examination ophthalmologic to establish the basal situation of the eye. The exam ophthalmologic includes indirect ophthalmoscopy, biomicroscopy with slit lamp, peripheral examination of the retina, measurement of the intraocular pressure, visual acuity (without correction and with maximum correction) and symptomatology.

After the preliminary examination, it is administered to patient an intrastromal injection of hyaluronidase ACS in the eye affected. If both eyes are affected, they can be treated separately. The stroma of the eye to be treated is injected with 40 µl of a composition containing 50 IU of the solution ACS hyaluronidase ophthalmic previously described to promote the reorganization of corneal collagen fibers and get the removal of the corneal scar over time.

After treatment, the eyes must be examined of the patient on days one (1), two (2), seven (7), fifteen (15), thirty (30) and sixty (60). On each day of the exam you will monitors the clarification of the corneal scar. After concluding The period of treatment the corneal scar has disappeared.

Example VII Use of hyaluronidase for the resolution of opacities corneal

In a patient who presents with an opacity corneal identifies the need for collagen reorganization corneal to improve your visual acuity. It is administered topically in the eye with opacity a pharmaceutical composition reorganizing the corneal collagen using a contact lens soft pretreated with the composition shown in Table IV. The corneal opacity is resolved as a result of treatment.

Example VIII Use of hyaluronidase to improve visual acuity after radial keratotomy (RK)

In a patient undergoing RK who presents with vision problems, including sparkles or star vision around the lights (cloud), the need for reorganization of corneal collagen fibers. During the RK procedure, the ophthalmologist makes a series of cuts (usually 4 to 8) in the cornea with a scalpel, following a pattern similar to the spokes of a wheel. These cuts are quite deep, sometimes affecting 90% of the thickness of the cornea. The cuts are similar to a "V" and make the central cornea relax or flatten and that the peripheral part of it becomes more  pending. These changes in the conformation of the cornea reduce the dome in its central part producing the improvement of vision not corrected After making the incisions the edges of them they will rejoin and healing will occur. During this process of healing the collagen fibers will fuse although the their organization in the area of incisions will not be the same quality as in the unaltered part of the cornea. Lack organization of collagen fibers in these areas with frequency results in corneal clouds.

About a week after RK procedure, the patient's solution is injected hyaluronidase presented in Table IV. Collagen fibers corneal areas of the incisions of the RK are reorganized as a consequence of the administration of hyaluronidase. The treatment result is the resolution of the cloud corneal

Example IX Use of hyaluronidase during radial keratotomy (RK) for improve visual acuity

To a patient who is going to have an RK as described in Example VIII the solution of Table IV hyaluronidase immediately after performing corneal incisions As a consequence of the administration of hyaluronidase, the collagen fibers that would heal the areas of incision made in the RK procedure, will be more organized that the fibers that have healed in the eye do not treaty. This procedure attached to the RK serves to prevent corneal clouds.

Example X Use of hyaluronidase to improve visual acuity after photorefractive keratotomy (PRK)

In a patient undergoing PRK who presents with vision defects, including sparkles or star vision around the lights (cloud) the need for reorganization of corneal collagen. Instead of making cuts in the cornea as in the RK, the PRK uses an excimer laser to Sculpt an area of 5 to 9 mm in diameter on the ocular surface. The process removes only 5-10% of the thickness of the cornea in mild myopia and up to 30% in myopia extreme - approximately the thickness of 1 to 3 human hairs. The main benefit of this procedure is that the integrity and resistance of the corneal dome. The laser excimeric is positioned at a wavelength of 193 mm, with that a corneal cell layer can be removed without damaging the adjacent cells. This allows the ophthalmologist to make modifications Extremely accurate cornea without injuring the eye.

When the cornea recovers from the procedure of PRK, the collagen fibers are repaired and regrouped. During this healing process undergoes the organization of collagen fibers resulting in the creation of corneal cloud.

About a week after PRK procedure, the patient is injected with the solution of hyaluronidase shown in Table IV. As a result of the administration of hyaluronidase collagen fibers are rearranged of the incision areas.

Example XI Use of hyaluronidase to improve visual acuity during photorefractive keratotomy (PRK)

To a patient undergoing PRK as described in Example X, the hyaluronidase solution is administered shown in Table IV in the form of drops, immediately after of laser treatment. Collagen fibers of the areas of incision made during the PRK procedure are rearranged as a consequence of the administration of hyaluronidase. The procedure attached to the PRK serves to prevent clouds corneal

Example XII Use of hyaluronidase to improve visual acuity after LASIK

In a patient undergoing LASIK and presenting with signs of defective vision, including flashes or vision of stars around the lights (cloud), the need for reorganization of corneal collagen fibers. Using a microkeratome or other similar device, the ophthalmologist "Lunch" the cornea from the side, producing a flap. A part of the device flattens the cornea during cutting, to Get a flap of uniform thickness. It is in this stage of procedure when the clinician has to have an extreme Caution and precision to create a perfect flap. After his creation, the flap folds back to expose the layers Corneal internal.

With the flap folded back the refractive correction of the inner layer of the cornea - which is performed with excimer laser similar to that used in the PRK. When have completed the treatment, the flap is placed back on its original position and the procedure ends. The eye has a natural suction facility that keeps the flap firmly in Your position at this time. The ophthalmologist should be careful to ensure an excellent fit when replacing the flap on your position As very little epithelium is altered, the Patients report a high level of comfort after the procedure. However, even when the flap has been perfectly relocated, there will be a repair process of collagen fibers corneal and some degree of disorganization on the periphery of the same.

Approximately one day after the procedure LASIK is injected into the patient with the hyaluronidase solution shown in Table IV. The corneal collagen fibers of the Incision areas in the LASIK procedure are reorganized as consequence of the administration of hyaluronidase. East procedure attached to the RK serves to prevent the cloud corneal

Example XIII Use of hyaluronidase during the LASIK procedure to improve visual acuity

To a patient undergoing a procedure of LASIK is given the hyaluronidase solution in Table IV, in the form of drops, immediately after remodeling the cornea, as described in Example XII. After the administration of the Hyaluronidase solution is placed on the flap and allowed to heal. After replacing the flap, the collagen fibers heal and grow in a more organized way compared to what happens in the corneal tissue treated with LASIK without applying the solution of hyaluronidase This procedure attached to LASIK serves to prevent corneal cloud.

References

Borcherding , M., Blacik , L., Sittg , A., Bizzell , J., Breen , M., and Weinstein , H. ( 1975 ). Proteoglycans and collagen fiber organization in human corneosileral tissue. Exp. Eye Res . 21, 59-70.

Campbell , C., Koester , C., Rittler , MC, Tackaberry , R ( 1974 ). Physiologic Optics, New York, NY: Harper &Row; 199-206.

Cintron , C., and Kublin , C. ( 1977 ) Dev. Biol . 61, 346-357.

Cintron , C., Gregory , JD, Dawle , PS, and Kublin , LC ( 1990 ). Invest. Ophthal & Vis. Sci. 31, 1975-1981.

Cintron , C., Hassinger , LC, Kublin , CL, and Cannon , DJ ( 1978 ). J. Ultrastract. Res. 65, 13-22.

Fitzsimmons , TD, Molander , N., Stenevi , U., Fagerholm , P., Schenholm , M., and Von Malmborg , A. ( 1994 ). Invest. Ophthalmol & Vis. Sci. 35, 2774-82.

Hassell , JR, Cintron , C., Kublin , CL, and Newsome , DA ( 1980 ). Proteoglycan changes during restoration of transparency in corneal scars. Archives of Biochemistry and Biophysics . 222 (2), 362-369.

Hassell , JR, Newsom , AD, Krachmer , JH, and Rodriguez , MM ( 1983 ). Proc. Natl Acad. Sci. 77, 3705-3709.

Howland , HC, Howland , B. ( 1977 ). A subjective method for the measurement of monochromatic aberrations of the eye. J. Opt. Soc. Am. 67, 1508-1518.

Kato , T., Nakayasu , K., Hosoda ., Y., Watanobe , Y., Kanai , A. ( 1999 ) Corneal wound healing following laser in situ keratomileuis (LASIK): a histopathological study in rabbits. J. Ophthalmol. 83, 1302-1305.

Maurice , DM ( 1957 ). The structure and transparency of the cornea. Br. J. Physiol. 136, 262-287.

Oshika , T., Klyce , S., Applegate , R., Howland , H., Danasoury , MA ( 1999 ). Comparison of corneal wavefront aberrations after photorefractive keratectomy and laser in situ keratomileusis. Am. J. of Ophthalmol. 127, 1-7.

Pallikaris , I. ( 1988 ). Quality of vision in refractive surgery. J. Refract. Surg. 14, 551-558.

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Schwarz , W., and Keyserlingk , DG ( 1969 ). In the cornea macromolecular organization of a connective tissue. John Hopkins Press, Baltimore / London. 123-132.

Toole , B., and Trelstad , R. ( 1971 ). Hyaluronate production and removal during corneal development in the chick. Dev. Biol. 26, 28-35.

Seiler , T., Holschback , F., Derse , M., Genth , U. ( 1994 ). Complications of myopic photorefractive keratectomy with the excimer laser. Ophthalmology 101, 153-160.

Seiler , T., Reckmann , W., Malong , RK ( 1993 ) Effective spherical aberration of the cornea as a quantitative descriptor in corneal topography. J. Cataract. Refract Surg. 19 (Suppl), 155-185.

Verdon , W., Bullimore , M., Maloney , RK ( 1996 ). Visual performance after photorefractive keratectomy: a prospective study. Arch. Ophthalmol. 114, 1465-1472.

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Claims (4)

1. Use of hyaluronidase free of fractions of molecular weight greater than 100,000, between 60,000 and 70,000 and per below 40,000, using 10% SDS-PAGE, for preparation of a medicine intended for the elimination of scars, opacities and corneal clouds. 2. Use according to claim 1, wherein said Hyaluronidase is administered in doses between 1 and 8,000 International Units 3. Use according to claims 1 or 2, in the that the medicine is intended for:

(to)

injection intrastromal,

(b)

application topical

(c)

administration through contact lenses,

(d)

administration unique and

(and)

 administration to At least twice.

4. Use according to any of the preceding claims, wherein said corneal opacity results from a surgical procedure selected from the group consisting of cataract surgery, corneal transplantation, radial keratotomy (RK), photorefractive keratectomy (PRK ) and keratomileusis in situ with laser (LASIK).